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采用腹腔注射α-半乳糖基神经酰胺(α-galactosylceramide,α-GC)的方法诱发C57BL/6小鼠流产,并检测注射后小鼠蜕膜NKT细胞表达穿孔素的情况,进而分析α-GC导致流产的发病机制。研究发现,在孕6.5 d腹腔注射α-GC(100μg/kg体重)可使小鼠孕10.5 d胚胎吸收率达到35.6%(37/104),显著高于对照组(7.4%;10/135;P<0.01)。此时采用流式细胞术检测蜕膜CD3+CD49b+NK1.1+细胞表达穿孔素的水平,发现α-GC刺激后穿孔素平均检出率显著高于对照组(15.5%和4.6%;P<0.01)。预先注射穿孔素的选择性抑制剂乙二醇四乙酸(ethylene glycol tetraacetic acid,EGTA)抑制穿孔素的作用,可显著降低α-GC所引起的小鼠胚胎吸收率增高。EGTA处理组和溶剂对照组α-GC诱导后小鼠胚胎吸收率分别为13.4%(16/119)和40.8%(42/103;P<0.01)。这些结果提示,α-GC可上调NKT细胞穿孔素的表达,而过多的穿孔素可能是α-GC导致小鼠胚胎吸收率增高的分子基础。
The abortion of C57BL / 6 mice was induced by intraperitoneal injection of α-galactosylceramide (α-GC), and the perforin NKT cells expressing perforin were detected after injection. The α-GC Cause the pathogenesis of abortion. The study found that intraperitoneal injection of α-GC (100 μg / kg body weight) at 6.5 d of gestation resulted in an embryo uptake rate of 35.6% (37/104) at 10.5 d of pregnancy, which was significantly higher than that of the control group (7.4%; 10/135; P <0.01). At this time, the level of perforin expression in decidual CD3 + CD49b + NK1.1 + cells was detected by flow cytometry and found that the average detection rate of perforin after α-GC stimulation was significantly higher than that of the control group (15.5% and 4.6%; P <0.01). Pretreatment of perforin with ethylene glycol tetraacetic acid (EGTA), a selective inhibitor of perforin, pretreatment with perforin significantly reduced the rate of embryo uptake by mice induced by α-GC. Absorption rates of embryos after induction by α-GC in EGTA-treated group and solvent-treated group were 13.4% (16/119) and 40.8% (42/103; P <0.01), respectively. These results suggest that α-GC can up-regulate the expression of perforin in NKT cells, while excessive perforin may be the molecular basis of α-GC in the induction of mouse embryo uptake.